A distal cis-regulatory element, CNS-9, controls NFAT1 and IRF4-mediated IL-10 gene activation in T helper cells
A distal cis-regulatory element, CNS-9, controls NFAT1 and IRF4-mediated IL-10 gene activation in T helper cells
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DOI：
10.1016/j.molimm.2008.07.037
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发表时间：
2009-02-01
影响因子：
3.6
通讯作者：
Im, Sin-Hyeog
中科院分区：
文献类型：
--
作者：
Lee, Choong-Gu;Kang, Kyu-Ho;Im, Sin-Hyeog
IL-10是一种多功能细胞因子，在维持免疫和耐受之间的平衡中起着关键作用。以前，我们确定了近端调控元件和染色质结构的改变，在IL-10基因位点的Th 1和Th 2细胞。我们现在已经确定了一个重要的顺式调控元件，CNS-9，位于9 kb的转录起始位点的IL-10基因位点的上游。CNS-9区域在脊椎动物基因组中高度保守，并且包含成簇的NFAT和IRF结合基序。通过EMSA观察到NFAT 1和IRF 4与CNS-9区域的体外结合。此外，通过ChIP观察到NFAT 1和IRF 4与CNS-9区域的Th 2优先体内结合。对野生型Th 2细胞或来自NFAT 1敲除（NFAT 1（-/-））小鼠的Th 2细胞的环孢霉素A处理显示CNS-9的反式活性显著降低。CNS-9的Th 2亚群特异性增强子活性通过NFAT 1及其伴侣IRF 4协同上调。NFAT 1和IRF 4结合位点的突变废除了CNS-9的增强子活性。总的来说，我们的研究结果确立了增强子元件CNS-9和与之结合的NFAT 1和IRF 4在不同辅助性T细胞亚群中IL-10表达的关键作用。(C)2008爱思唯尔有限公司保留所有权利。
IL-10 is a multifunctional cytokine that plays a critical role in maintaining the balance between immunity and tolerance. Previously, we identified proximal regulatory elements and alterations of chromatin structure in the IL-10 gene loci of Th1 and Th2 cells. We have now characterized a crucial cis-regulatory element, CNS-9, located 9 kb upstream of the transcription start site in IL-10 gene loci. The CNS-9 region is highly conserved in vertebrate genomes, and contains clustered NFAT and IRF binding motifs. In vitro binding of NFAT1 and IRF4 to the CNS-9 region was observed by EMSA. Furthermore, Th2-preferential in vivo binding of NFAT1 and IRF4 to the CNS-9 region was observed by ChIP. Cyclosporine A treatment on wild type Th2 cells or Th2 cells derived from NFAT1 knockout (NFAT1(-/-)) mice showed significantly reduced trans-activity of CNS-9. The Th2 subset-specific enhancer activity of CNS-9 was upregulated synergistically by NFAT1 and its partner IRF4. Mutations in the binding sites for NFAT1 and IRF4 abrogated its enhancer activity of CNS-9. Collectively, our results establish crucial roles for enhancer element CNS-9, and NFAT1 and IRF4 that bind to it, for IL-10 expression in differential T helper subsets. (C) 2008 Elsevier Ltd. All rights reserved.